A mixture of biotinylated monoclonal antibodies against the non-CD8+ T cells is added to the starting sample and allowed to bind to the cells. Depletion MyOne™ SA Dynabeads® are added and will bind to the antibody-labelled cells during a short incubation. The bead-bound cells are subsequently separated on a magnet and discarded. The remaining, untouched human CD8+ T cells can be used for any application.

* Note that the difference in volumes for small vs. large scale protocol is based on getting good working volumes during incubation with beads. A shift from small to large scale parameters may be done at any level of cells processed, as long as the resulting volume during incubation with beads is appropriate for the tubes used (more than 1/10 of total tube volume).

The starting number of cells for this protocol is 5 x 107 (see preparations above for details)

Transfer 500 μl (5 × 107) PBMC in Isolation Buffer to a tube.

Add 100 μl heat inactivated FCS.

Add 100 μl of Antibody Mix.

Mix well and incubate for 20 min at 2-8°C.

Wash the cells by adding 10 ml Isolation Buffer. Mix well by tilting the tube several times and centrifuge at 350 × g for 8 min at 2-8°C. Discard the supernatant.

Resuspend the cells in 500 μl Isolation Buffer.

Add 500 μl pre-washed Depletion MyOne SA Dynabeads®.

Incubate for 15 min at 18-25°C with gentle tilting and rotation.

Resuspend the bead-bound cells by vigorously pipetting >10 times using a pipette with a narrow tip opening (e.g. a 1000 μl pipette tip or a 5 ml serological pipette).

General Information

Invitrogen Dynal® AS complies with the Quality System Standards ISO 9001:2000 and ISO 13485:2003.

Storage/Stability

This product is stable until the expiry date stated on the label when stored unopened at 2-8°C. Store opened vials at 2-8°C and avoid bacterial contamination. Keep Dynabeads® in liquid suspension during storage and all handling steps, as drying will result in reduced performance. Resuspend well before use.

Warnings And Limitations

This product is for research use only. Not intended for any animal or human therapeutic or diagnostic use unless otherwise stated. Follow appropriate laboratory guidelines. This product contains 0.02% sodium azide as a preservative, which is cytotoxic.

Avoid pipetting by mouth!

Sodium azide may react with lead and copper plumbing to form highly explosive metal azides. When disposing through plumbing drains, flush with large volumes of water to prevent azide build up. Certificate of Analysis (CoA) is available upon request. Material Safety Data Sheet (MSDS) is available at
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